To study the chemical constituents of Asarum himalaicum, fifteen compounds were isolated from a 70% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, and HR-ESI-MS, these compounds were identified as 4-demethoxyaristolochic acid BII (1), aristolochic acid I (2), aristolochic acid Ia (3), 7-hydroxyaristolochic acid I (4), aristolochic acid IV (5), aristolic acid II (6), debilic acid (7), aristololactam I (8), 9-hydroxyaristololactam I (9), 7-methoxyaristololactam IV (10), (2S)-narigenin-5, 7-di-O-beta-D-pyranosylglucoside (11), 4-hydroxybenzoic acid (12), 3, 4-dihydroxybenzoic acid (13), 4-hydroxycinnamic acid (14), and beta-sitosterol (15). All of these compounds (1-15) were obtained from A. himalaicum for the first time. Among them, 1 was identified as a new compound, and compounds 3-6, 9, 12-14 were isolated from Asarum genus for the first time. Since the kidney toxicity of aristolochic acids and aristololactams has been reported, the result of this investigation suggests that it should be cautioned to use A. himalaicum as a medicine.

The blood concentrations of the pharmacodynamic substances of traditional Chinese medicines (TCMs) are usually very low. How can they exert pharmacological actions, in which forms (original form, metabolite or the both) do they exert the actions. To answer these questions, we proposed a new concept ofEffective Formsof pharmacodynamic substances of TCMs and a hypothesis of additive effect of multiple constituents of TCMs. The hypothesis includes that the aggregate or summation of Effective Forms of pharmacodynamic substances of TCMs is the core material base of the effi-cacy of TCMs, and the additive effect of the blood concentrations of different Effective Forms is one part of the action mechanism. The additive effect of the different Effective Forms of a TCMs means an additive effect of numerous con-stituents or/and metabolites on a same target, and therefore the efficacy brought by the addition of the concentrations of all these compounds, which different from the synergy effect of multi-constituents on multi-targets. Studies on the disposition of TCMs showed that a constituent can be biotransformed to many metabolites (up to more than 50 metabolites);different constituents can produce the same metabolites;many metabolites (up to 10 compounds for each metabolite) are isomers or homologues; some constituents can be converted to each other in vivo; and some metabolites are bioactive. These com-pounds having the similar structure are likely to have the same pharmacological effects on the same target, which could provide experimental evidences for the concept ofEffective Formsand the hypothesis ofAdditive Effect. We suggest that the Effective Forms and Additive Effects of the pharmacodynamic substances of TCMs should be extensively investi-gated in the future, and the results of such researches will help us further understand the pharmacodynamic substances and action mechanism of TCMs, and give a new explanation 'Toxicities Scattering Effect' for 'Why the toxicities of TCMs are low', and propose a new strategy for quality control of TCMs.

OBJECTIVE:To establish the HPLC characteristic chromatogram of pheretima,and compare the differences of the main ingredient contents of Guangdong pheretima and Shanghai pheretima and the chromatogram differences among pheretima and 3 other animal drugs. METHODS:Pheretima HPLC characteristic chromatogram method was adopted to determine the characteris-tic chromatograms of 16 Guangdong pheretima,8 Shanghai pheretima,3 eupolyphaga,3 hirudo and 3 catharsius. Similarity evalua-tion and t test were used to analyze the differences of chromatogram data of 5 animal drugs. RESULTS:The established HPLC char-acteristic chromatogram method firstly identified 11 common characteristic peaks,including 6 nucleosides,4 nucleobase and 1 ami-no acid;and it could be used for the identification of pheretima from eupolyphaga,hirudo and catharsius;the differences of main ingredient contents in the characteristic chromatogram of Guangdong pheretima and Shanghai pheretima were firstly studied. The contents of xanthine and adenosine in Guangdong pheretima were higher than Shanghai pheretima,while the contents of uridine, guanosine and 2′-deoxy guanosine in Shanghai pheretima were higher than Guangdong pheretima. A new index S,calculated by these 5 constituents,was successfully applied to distinguish the 2 kinds of pheretima. CONCLUSIONS:The characteristic chro-matogram can be used for the identification of pheretima,and can provide reference for the pharmacodynamic differences study of Guangdong pheretima and Shanghai pheretima.

OBJECTIVE:To establish a method of simultaneous determination of inulicin and deacetylinulicin in Inulae Flos. METHODS:HPLC was performed on the column of Zorbax SB-C18 with mobile phase of acetonitrile-water(gradient elution)at a flow rate of 1.0 ml/min,the column temperature was 25 ℃,the detection wavelength was 210 nm,and the injection volume was 10 μl. RESULTS:The linear range was 0.000 2-0.005 μg/ml(r=0.999 8)for inulicin and 0.000 1-0.001 7 μg/ml(r=0.999 4)for deacetylinulicin;RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries were 99.63%-103.56%(RSD=1.26%,n=9)and 95.98%-101.21%(RSD=1.84%,n=9),respectively. CONCLUSIONS:The method is simple,accurate and reliable,and can be used for the quality evaluation of Inulae Flos.

OBJECTIVE:To study the metabolites,distribution,metabolic type and the possible activity of harpagide which is the active component from Scrophularia ningpoensis in rats in vivo. METHODS:4 SD rats were divided into blank group (ul-tra-pure water) and administration group (harpagide reference solution),2 in each group,ig,160 mg/kg,twice a day,for 3 d. Urine and feces were collected every 12 h before administration and the first administration;sample blood 8 mL was taken after 0.5,1 h of last administration;heart,liver,spleen,lung,kidney,stomach and small intestine were taken. The blood,urine,fe-ces and other tissue solutions were prepared,HPLC-MS was conducted to detect and identify the harpagide metabolites in rats in vi-vo and presume metabolic pathways,and PharmMapper software was used to predict metabolites activity. RESULTS:12 harpagide metabolites were identified in rats in vivo,the form of prototypes and metabolites were distributed in heart,liver,spleen,lung, kidney,stomach and small intestine. The metabolic type mainly included hydrolysis,dehydration,reduction,methylation,sul-fation,glucuronic acid binding,grade A coumaric acid binding,etc. The 12 compounds may have activities in the treatment of epi-lepsy,amyotrophic lateral sclerosis,diabetes,stroke,etc. CONCLUSIONS:Harpagide may be effective in the form of prototypes and metabolites. The study has provided basis for attributing the origins of metabolite,studying the effective form of S. ningpoensis clarifying its pharmacological mechanism and processing mechanism.

OBJECTIVE:To explore the effects and mechanism of extracts,active constituents and constituent combination of Sinopodophylli Fructus on cell proliferation of human breast cancer. METHODS:Acid phosphatase method was conducted to deter-mine the effects of 4 extracts [ethanol extract (Xc),petroleum ether extract from ethanol extract (Xp),ethyl acetate extract from ethanol extract (Xe),n-butanol extract from ethanol extract (Xz)],5 active constituents [podophyllotoxin (S1),deoxypodophyllo-toxin (S2),4-desmethyl deoxypodophyllotoxin (S3),8-isopentenyl kaempferol (S4),8,2′-diisoprenyl quercetin-3-methyl ether (S5)] and 3 active constituent combination [combination 1,S1-S2-S3-S4-S5 (2:4:1:4:32),Z1;combination 2,S2-S4 (1:1),Z2;combination 3,S3-S4(1:4),Z3] on the MDA-MB-231,MCF-7 cell proliferation;flow cytometry was adopted to detect the effects of above-mentioned samples on MDA-MB-231,MCF-7(T47D)cell cycle and mitochondrial membrane potential. RESULTS:The active constituent combination Z1 showed significant inhibitory effects on MDA-MB-231,MCF-7 cells,the half inhibitory concen-trations(IC50)were(0.27±0.2),(0.11±0.1)μg/mL;extracts Xc,Xp,Xe,active constituents S2,S4 and active constituent combi-nation Z2,Z3 showed relatively strong inhibitory effects on MDA-MB-231,MCF-7 (T47D) cell proliferation (IC50<15 μg/mL). Both extracts and active constituents can block MDA-MB-231,MCF-7 cell cycle in G2/M phase;all active constituents can block MDA-MB-231,T47D cell cycle in G0/G1 phase,and can reduce MDA-MB-231,T47D cell mitochondrial membrane potential. CONCLUSIONS:The active constituents and constituent combination of Sinopodophylli Fructus can inhibit cell proliferation of breast cancer by affecting cell cycle and mitochondrial mem-brane potential.

OBJECTIVE:To establish a method for the contents determination of 9 components in Guizhi decoction,and com-pare the effects of traditional decoction method and the extracting machine decoction method on these contents in Guizhi decoction. METHODS:HPLC was performed on the column of Agilent Zorbax SB-C18 with mobile phaseof acetonitrile- 0.1% phosphoric ac-id(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 230 nm,254 nm and 280 nm,the column tempera-ture was 25℃,and the injection volume was 10μl. RESULTS:The linear range was 0.410 2-210.0μg/ml for gallic acid(r=0.999 9), 0.994 0-254.5μg/ml for albiflorin(r=0.999 9),1.636 0-1 675.0μg/ml for paeoniflorin(r=0.999 9),0.988 3-506.0μg/ml for liquiri-tin(r=0.999 6),0.987 3-31.59 μg/ml for coumarin(r=0.999 5),0.486 8-124.6 μg/ml for cinnamic acid(r=0.999 5),2.458 0-314.6μg/ml for cinnamaldehyde(r=0.999 5),0.034 3-1.096 μg/ml for 2-methoxy cinnamaldehyde(r=0.999 8),and 1.711 0-219.0 μg/ml for glycyrrdhizic acid (r=0.999 7);RSDs of precision,stability and reproducibility tests were lower than 5%,recoveries were 93.56%-103.19%(RSD=4.00%,n=9)、101.51%-107.32%(RSD=2.21%,n=9)、95.08%-103.76%(RSD=2.87%,n=9)、100.82%-105.73%(RSD=1.85%,n=9)、85.08%-89.12%(RSD=1.40%,n=9)、92.31%-99.12%(RSD=2.71%,n=9)、99.17%-102.32%(RSD=1.24%,n=9)、100.15%-103.98%(RSD=1.18%,n=9)、99.93%-102.61%(RSD=1.03%,n=9). The content of total effective components from the extracting machine decoction method was 4 565μg/g,that from the traditional decoc-tion method was 2 742 μg/g.CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of 9 componentsin Guizhi decoction. The contents of gallic acid,albiflorin and 2-methoxy cinnamaldehyde are first re-ported. The total effective components from the extracting machine decoction method are higher than that from the traditional decoc-tion method.

Objective To investigate the in vitro effects of various antibiotics (spectinomycin, ceftriaxone, erythromycin, ofloxacin and doxycycline) against 12 isolates of C. trachomatis. Methods Minimal inhibitory concentrations (MICs ) and fractional inhibitory concentrations (FICs) of the antimicrobials against all C. trachomatis were calculated. Checkerboard method was used for the determination of FICs and Ridit test for the comparison of the interactions among the various combinations. Results No difference was observed in most of the combinations. No antagonism was found in all except for ceftriaxone-doxycycline combination. Synergism was observed in 42% (5 of 12) and 50% (6 of 12) of the chlamydial isolates for erythromycin-spectinomycin and doxycycline-spectinomycin combination, respectively. No significant difference was observed among triple combinations with spectinomycin or with ceftriaxone. When interactions of erythromycin, ofloxacin and doxycycline with spectinomycin were compared to those with ceftriaxone respectively, both interactions of erythromycin (U = 2.46, P = 0.014) and doxycycline (U = 2.83, P = 0.002) were more synergistic with spectinomycin than those with ceftriaxone. Conclusions This study indicates that the combination of spectinomycin with erythromycin or doxycycline is more effective against C. trachomatis than that of ceftriaxone. Therefore, spectinomycin rather than ceftriaxone might be recommended in the dual therapy against C. trachomatis and N. gonorrhoeae.

Object To study the antitumor effects of extracts from the fruits, roots and rhizomes of Sinopodophyllum emodi (Wall.) Ying, the roots and rhizomes of Dysosma furfuracea S.Y. Bao and the podophyllotoxin, deoxypodophyllotoxin of Guijiu plants in mice. Methods Tumor borne mice with transplanted HepA and Ehrlich ascites carcinoma (EAC) were used to observe the antitumor and the liver protective effects of the above five samples, which were given orally. Results Extracts of roots and rhizomes of S. emodi, and D. furfuracea showed the significant antitumor effects. Conclusion All the five samples have antitumor activity.

OBJECTIVETo study the chemical constituents of the fruit of Aristolochia contorta.

METHODThe compounds were isolated by chromatographic techniques and crystalization, the structures were elucidated by spectrum analysis.

RESULTFifteen compounds were isolated from the dry fruit of A. contorta, which were six aristolochic acids: aristolochic acid I, aristolochic acid III a, aristolochic acid IVa, aristolochic acid II, aristolochic acid III and aristolochic acid VIIa. Three aristolactams: aristololactam I, aristololactam II and aristololactam IIIa. Three phenolic acids syringic acid, vanillic acid and p-coumaric acid. Three other type compounds: pentacosane acid, beta-sitosterol and daucossterol.

CONCLUSIONAristolochic acid III, aristolochic acid VIIa, aristololactam IIIa, and penfacosane acid were isolated from A. contorta for the first time, and compounds 4-13 were isolated from the furit of A. contorta for the first time.